Cell Type-specific Translational Profiling in the Xenopus laevis Retina
| Autor: | Elizabeth A. Mills, X. Wang, F. L. Watson, D. F. Chen, C. Guo, Nicholas Marsh-Armstrong |
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| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Retinal Ganglion Cells
Cell Xenopus Xenopus Proteins Real-Time Polymerase Chain Reaction Ribosome Retinal ganglion Article Xenopus laevis Ribosomal protein Protein biosynthesis medicine Animals RNA Messenger Eye Proteins Retina biology Gene Expression Profiling biology.organism_classification Molecular biology Gene expression profiling medicine.anatomical_structure Gene Expression Regulation Protein Biosynthesis Developmental Biology Photoreceptor Cells Vertebrate |
| Popis: | Background: Translating Ribosome Affinity Purification (TRAP), a method recently developed to generate cell type–specific translational profiles, relies on creating transgenic lines of animals in which a tagged ribosomal protein is placed under regulatory control of a cell type–specific promoter. An antibody is then used to affinity purify the tagged ribosomes so that cell type–specific mRNAs can be isolated from whole tissue lysates. Results: Here, cell type–specific transgenic lines were generated to enable TRAP studies for retinal ganglion cells and rod photoreceptors in the Xenopus laevis retina. Using real time quantitative PCR for assessing expression levels of cell type–specific mRNAs, the TRAP method was shown to selectively isolate mRNAs expressed in the targeted cell and was efficient at purifying mRNAs expressed at both high and low levels. Statistical measures used to distinguish cell type–specific RNAs from low level background and non-specific RNAs showed TRAP to be highly effective in Xenopus. Conclusions: TRAP can be used to purify mRNAs expressed in rod photoreceptors and retinal ganglion cells in X. laevis. The generated transgenic lines will enable numerous studies into the development, disease, and injury of the X. laevis retina. Developmental Dynamics, 2012. © 2012 Wiley Periodicals, Inc. |
| Databáze: | OpenAIRE |
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